Notes

  • Unclassified reads were excluded
  • Taxa with total reads number of 10 or lower were excluded
  • Eukaryotes were excluded from the analyis

Import Exp data

MCM D6300

MCM D6331

FR D6323

Canis lupus

Homo sapiens

Merge

Filter Exp data

Filter samples based on total read count

Fix sample names

Add lineage and metadata to Gold Standard Data

Limosilactobacillus fermentum in the theoretical composition was changed to Lactobacillus fermentum

## Error in validObject(.Object) : invalid class "otu_table" object: 
##  OTU abundance data must have non-zero dimensions.

Fix sample names from ‘-’

Merge exp data with GS

Filter Taxa for Gold Standard

These are the taxa that were found in the experimental data

Add Gram Stain Info

Omit Eukaryotes

Zymo ONT V1-V9 comparison: Relative Abundance of Taxa at each Taxonomic Level

Parameters and Settings

Correlation

PERMANOVA model

Gram Stain level

Barplot of abundances

Phylum level

Barplot of abundacnes

MAASLIN2 - Number of Significantly Different Taxa Among FR, H. sapiens and C. lupus

Barplot of Coefficients

Heatmap of Significant Associations (−log(qval) x sign(coeff))

PERMANOVA

## Permutation test for adonis under reduced model
## Terms added sequentially (first to last)
## Blocks:  strata 
## Permutation: free
## Number of permutations: 999
## 
## adonis2(formula = otutab_t ~ primer, data = sampdat, method = distance, by = "terms", strata = sampdat$source)
##          Df SumOfSqs      R2      F Pr(>F)
## primer    2   0.0964 0.01882 0.2301  0.367
## Residual 24   5.0274 0.98118              
## Total    26   5.1238 1.00000

Class level

Barplot of abundances

MAASLIN2 - Number of Significantly Different Taxa Among FR, H. sapiens and C. lupus

Barplot of Coefficients

Heatmap of Significant Associations (−log(qval) x sign(coeff))

PERMANOVA

## Permutation test for adonis under reduced model
## Terms added sequentially (first to last)
## Blocks:  strata 
## Permutation: free
## Number of permutations: 999
## 
## adonis2(formula = otutab_t ~ primer, data = sampdat, method = distance, by = "terms", strata = sampdat$source)
##          Df SumOfSqs     R2      F Pr(>F)
## primer    2    0.136 0.0252 0.3102  0.175
## Residual 24    5.261 0.9748              
## Total    26    5.397 1.0000

Order level

Barplot of abundances

MAASLIN2 - Number of Significantly Different Taxa Among FR, H. sapiens and C. lupus

Barplot of Coefficients

Heatmap of Significant Associations (−log(qval) x sign(coeff))

### PERMANOVA

## Permutation test for adonis under reduced model
## Terms added sequentially (first to last)
## Blocks:  strata 
## Permutation: free
## Number of permutations: 999
## 
## adonis2(formula = otutab_t ~ primer, data = sampdat, method = distance, by = "terms", strata = sampdat$source)
##          Df SumOfSqs      R2      F Pr(>F)
## primer    2   0.1361 0.02519 0.3101  0.204
## Residual 24   5.2675 0.97481              
## Total    26   5.4036 1.00000

Family level

Barplot of abundances

MAASLIN2 - Number of Significantly Different Taxa Among FR, H. sapiens and C. lupus

Barplot of Coefficients

Heatmap of Significant Associations (−log(qval) x sign(coeff))

PERMANOVA

## Permutation test for adonis under reduced model
## Terms added sequentially (first to last)
## Blocks:  strata 
## Permutation: free
## Number of permutations: 999
## 
## adonis2(formula = otutab_t ~ primer, data = sampdat, method = distance, by = "terms", strata = sampdat$source)
##          Df SumOfSqs      R2      F Pr(>F)  
## primer    2   0.1690 0.02807 0.3466  0.075 .
## Residual 24   5.8511 0.97193                
## Total    26   6.0202 1.00000                
## ---
## Signif. codes:  0 '***' 0.001 '**' 0.01 '*' 0.05 '.' 0.1 ' ' 1

Genus level

Barplot of abundances

MAASLIN2 - Number of Significantly Different Taxa Among FR, H. sapiens and C. lupus

Barplot of Coefficients

Heatmap of Significant Associations (−log(qval) x sign(coeff))

PERMANOVA

## Permutation test for adonis under reduced model
## Terms added sequentially (first to last)
## Blocks:  strata 
## Permutation: free
## Number of permutations: 999
## 
## adonis2(formula = otutab_t ~ primer, data = sampdat, method = distance, by = "terms", strata = sampdat$source)
##          Df SumOfSqs      R2      F Pr(>F)
## primer    2   0.1383 0.02149 0.2635  0.163
## Residual 24   6.2977 0.97851              
## Total    26   6.4360 1.00000

Species level

Barplot of abundances

MAASLIN2 - Number of Significantly Different Taxa Among FR, H. sapiens and C. lupus

Barplot of Coefficients

Heatmap of Significant Associations (−log(qval) x sign(coeff))

PERMANOVA

## Permutation test for adonis under reduced model
## Terms added sequentially (first to last)
## Blocks:  strata 
## Permutation: free
## Number of permutations: 999
## 
## adonis2(formula = otutab_t ~ primer, data = sampdat, method = distance, by = "terms", strata = sampdat$source)
##          Df SumOfSqs      R2     F Pr(>F)  
## primer    2   0.1743 0.02423 0.298  0.038 *
## Residual 24   7.0199 0.97577               
## Total    26   7.1942 1.00000               
## ---
## Signif. codes:  0 '***' 0.001 '**' 0.01 '*' 0.05 '.' 0.1 ' ' 1

Summary of Similarity of FR Taxa to Homo sapiens and Canis lupus

Summary of PERMANOVAs on Different Rank Levels

Summary of Detection and Correlation Statistics for Zymo MCM D6300, D6331 and D6323

Detection Statistics

##                                  sample     N
##                                  <char> <int>
## 1: PRMT_FM1_FR_MONT_20240807_bc25_D6323    60
## 2: PRMT_FM2_FR_MONT_20240807_bc26_D6323    60
## 3: PRMT_FM3_FR_MONT_20240807_bc27_D6323    60
## 4:  PRMT_FO1_FR_ONT_20240807_bc10_D6323    60
## 5:  PRMT_FO2_FR_ONT_20240807_bc11_D6323    60
## 6:  PRMT_FO3_FR_ONT_20240807_bc12_D6323    60
## 7:   PRMT_FP1_FR_PB_20240807_bc40_D6323    60
## 8:   PRMT_FP2_FR_PB_20240807_bc41_D6323    60
## 9:   PRMT_FP3_FR_PB_20240807_bc42_D6323    60
##                                   sample     N
##                                   <char> <int>
## 1: PRMT_MM1_MCS_MONT_20240807_bc19_D6300    60
## 2: PRMT_MM2_MCS_MONT_20240807_bc20_D6300    60
## 3: PRMT_MM3_MCS_MONT_20240807_bc21_D6300    60
## 4:  PRMT_MO1_MCS_ONT_20240807_bc04_D6300    60
## 5:  PRMT_MO2_MCS_ONT_20240807_bc05_D6300    60
## 6:  PRMT_MO3_MCS_ONT_20240807_bc06_D6300    60
## 7:   PRMT_MP1_MCS_PB_20240807_bc34_D6300    60
## 8:   PRMT_MP2_MCS_PB_20240807_bc35_D6300    60
## 9:   PRMT_MP3_MCS_PB_20240807_bc36_D6300    60
##                                   sample     N
##                                   <char> <int>
## 1: PRMT_GM1_GMS_MONT_20240807_bc22_D6331    60
## 2: PRMT_GM2_GMS_MONT_20240807_bc23_D6331    60
## 3: PRMT_GM3_GMS_MONT_20240807_bc24_D6331    60
## 4:  PRMT_GO1_GMS_ONT_20240807_bc07_D6331    60
## 5:  PRMT_GO2_GMS_ONT_20240807_bc08_D6331    60
## 6:  PRMT_GO3_GMS_ONT_20240807_bc09_D6331    60
## 7:   PRMT_GP1_GMS_PB_20240807_bc37_D6331    60
## 8:   PRMT_GP2_GMS_PB_20240807_bc38_D6331    60
## 9:   PRMT_GP3_GMS_PB_20240807_bc39_D6331    60
##              statistic     N
##                 <char> <int>
##  1:     True_positives   162
##  2:    False_positives   162
##  3:    False_negatives   162
##  4:          Precision   162
##  5:             Recall   162
##  6:                 F1   162
##  7:               F0.5   162
##  8:           Chi_stat   162
##  9: Chisq.test.p_value   162
## 10:    Chisq.test.sign   162
##    GS_version     N
##        <fctr> <int>
## 1:      D6331   540
## 2:      D6300   540
## 3:      D6323   540

Chi^2

F1, Precision and Recall

Correlation of Abundance Values between Theoretical and Experimental

##                                  sample     N
##                                  <char> <int>
## 1: PRMT_FM1_FR_MONT_20240807_bc25_D6323     4
## 2: PRMT_FM2_FR_MONT_20240807_bc26_D6323     4
## 3: PRMT_FM3_FR_MONT_20240807_bc27_D6323     4
## 4:  PRMT_FO1_FR_ONT_20240807_bc10_D6323     4
## 5:  PRMT_FO2_FR_ONT_20240807_bc11_D6323     4
## 6:  PRMT_FO3_FR_ONT_20240807_bc12_D6323     4
## 7:   PRMT_FP1_FR_PB_20240807_bc40_D6323     4
## 8:   PRMT_FP2_FR_PB_20240807_bc41_D6323     4
## 9:   PRMT_FP3_FR_PB_20240807_bc42_D6323     4
##                                   sample     N
##                                   <char> <int>
## 1: PRMT_MM1_MCS_MONT_20240807_bc19_D6300     4
## 2: PRMT_MM2_MCS_MONT_20240807_bc20_D6300     4
## 3: PRMT_MM3_MCS_MONT_20240807_bc21_D6300     4
## 4:  PRMT_MO1_MCS_ONT_20240807_bc04_D6300     4
## 5:  PRMT_MO2_MCS_ONT_20240807_bc05_D6300     4
## 6:  PRMT_MO3_MCS_ONT_20240807_bc06_D6300     4
## 7:   PRMT_MP1_MCS_PB_20240807_bc34_D6300     4
## 8:   PRMT_MP2_MCS_PB_20240807_bc35_D6300     4
## 9:   PRMT_MP3_MCS_PB_20240807_bc36_D6300     4
##                                   sample     N
##                                   <char> <int>
## 1: PRMT_GM1_GMS_MONT_20240807_bc22_D6331     4
## 2: PRMT_GM2_GMS_MONT_20240807_bc23_D6331     4
## 3: PRMT_GM3_GMS_MONT_20240807_bc24_D6331     4
## 4:  PRMT_GO1_GMS_ONT_20240807_bc07_D6331     4
## 5:  PRMT_GO2_GMS_ONT_20240807_bc08_D6331     4
## 6:  PRMT_GO3_GMS_ONT_20240807_bc09_D6331     4
## 7:   PRMT_GP1_GMS_PB_20240807_bc37_D6331     4
## 8:   PRMT_GP2_GMS_PB_20240807_bc38_D6331     4
## 9:   PRMT_GP3_GMS_PB_20240807_bc39_D6331     4

R^2 Barplot

R^2 Lineplot

B-diversity Analysis using sample-wise Bray-Curtis distances

Non-metric Multi-Dimensional Scaling (NMDS)

NMDS v1: color = DNA_isolation_method; shape = Vregion

NMDS v2: color = Vregion; shape = DNA_isolation_method